分析化学2013，Vol.41Issue(7)：1025-1030,6.DOI:10.3724/SP.J.1096.2013.20972
胶体金免疫层析法快速检测牛奶、奶粉、饲料中的三聚氰胺
A Colloidal Gold Immunochromatographic Assay for Rapid Detection of Melamine in Milk, Milk Powder and Animal Feeds
摘要
Abstract
A colloidal gold immunochromatographic strip for rapid detection of melamine in milk,milk powder and animal feeds was presented.The molecular structure of melamine was rationally modified.Two melamine derivatives were synthesized and coupled to bovine serum albumin (BSA) and ovalbumin (OVA) for the production of immunogen and coating antigen.Monoclonal antibody (mAb) against melamine was produced by hybridoma technology.Colloidal gold nanoparticles with the average particle diameter of 18 nm were linked with mAb to form the Au-mAb conjugate.The formed Au-mAb was coated on glass fiber; while the coating antigen and goat anti-mouse IgG were separately sprayed onto the nitrocellulose (NC) membrane as the test (T) line and the control (C) line.The Au-mAb pad,NC membrane,sample pad and absorptive paper were assembled into an immunochromatographic strip.Standard (or sample) solution was added to the sample pad.After 10 min,the colloidal gold color (red) was visually observed at C line and T line,and the content of the melamine was evaluated by comparing the intensity of the color on these two lines.The results demonstrated that the limit of detection (LOD) of the melamine was 10 μg/L.Ten compounds were selected for specificity testing.The cross-reactivity (CR) of the immunochromatographic strip with 2-chloro-4,6-diamino-1,3,5-triazine (CAAT) and cyromazine was about 1％,and there was no CR of the strip with other eight compounds.The samples spiked with melamine at different concentrations were detected by immunochromatographic assay and enzyme-linked immunosorbent assay (ELISA) simultaneously.The results obtained by these two methods were in a good agreement.The proposed immunochromatographic assay can be utilized as a promising tool for rapid detection of melamine in milk,milk powder and feed samples.关键词
三聚氰胺/单克隆抗体/胶体金/免疫层析法Key words
Melamine / Monoclonal antibody / Colloidal gold / Immunochromatographic assay引用本文复制引用
李淑群,曹碧云,常化仿,邓安平..胶体金免疫层析法快速检测牛奶、奶粉、饲料中的三聚氰胺[J].分析化学,2013,41(7):1025-1030,6.基金项目
本文系国家自然科学基金(No.21175097)资助 （No.21175097）
